Literature DB >> 36173567

Low-Input CUT&RUN for Mouse Oocytes and Preimplantation Embryos.

Ryoya Hayashi1,2, Azusa Inoue3,4.   

Abstract

Cleavage Under Target & Release Using Nuclease (CUT&RUN) enables the detection of DNA regions that are bound by a protein of interest. This method is suitable for low-input materials because of the absence of an immunoprecipitation step. However, it sometimes fails when applying it to fragile cells, such as mouse oocytes. Here we describe our low-input CUT&RUN protocol optimized for mouse oocyte and preimplantation embryo samples in which the primary antibody and protein A-MNase binding steps are completed before the cells are bound to Concanavalin A-coated magnetic beads. This modification prevents crush of oocytes and early embryos and unwanted loss of chromatin during CUT&RUN procedures.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Entities:  

Keywords:  Epigenetics; Histone modification; Oocytes; Preimplantation embryo

Mesh:

Substances:

Year:  2023        PMID: 36173567     DOI: 10.1007/978-1-0716-2724-2_6

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  6 in total

1.  Resetting Epigenetic Memory by Reprogramming of Histone Modifications in Mammals.

Authors:  Hui Zheng; Bo Huang; Bingjie Zhang; Yunlong Xiang; Zhenhai Du; Qianhua Xu; Yuanyuan Li; Qiujun Wang; Jing Ma; Xu Peng; Feng Xu; Wei Xie
Journal:  Mol Cell       Date:  2016-09-15       Impact factor: 17.970

2.  H2AK119ub1 guides maternal inheritance and zygotic deposition of H3K27me3 in mouse embryos.

Authors:  Hailiang Mei; Azusa Inoue; Chisayo Kozuka; Ryoya Hayashi; Mami Kumon; Haruhiko Koseki
Journal:  Nat Genet       Date:  2021-04-05       Impact factor: 38.330

3.  An efficient targeted nuclease strategy for high-resolution mapping of DNA binding sites.

Authors:  Peter J Skene; Steven Henikoff
Journal:  Elife       Date:  2017-01-16       Impact factor: 8.140

4.  Genomic imprinting of Xist by maternal H3K27me3.

Authors:  Azusa Inoue; Lan Jiang; Falong Lu; Yi Zhang
Journal:  Genes Dev       Date:  2017-10-01       Impact factor: 11.361

5.  Maternal Eed knockout causes loss of H3K27me3 imprinting and random X inactivation in the extraembryonic cells.

Authors:  Azusa Inoue; Zhiyuan Chen; Qiangzong Yin; Yi Zhang
Journal:  Genes Dev       Date:  2018-11-21       Impact factor: 11.361

6.  Improved CUT&RUN chromatin profiling tools.

Authors:  Michael P Meers; Terri D Bryson; Jorja G Henikoff; Steven Henikoff
Journal:  Elife       Date:  2019-06-24       Impact factor: 8.140

  6 in total

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